LOCUS       sequence-92160-e        4893 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MethyloBrick cloning vector, no promoter; KmR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4893)
  AUTHORS   Schada von Borzyskowski L, Remus-Emsermann M, Weishaupt R, Vorholt 
            JA, Erb TJ
  TITLE     A Set of Versatile Brick Vectors and Promoters for the Assembly, 
            Expression, and Integration of Synthetic Operons in Methylobacterium
            extorquens AM1 and Other Alphaproteobacteria.
  JOURNAL   ACS Synth Biol. 2014 Aug 8.
  PUBMED    25105793
REFERENCE   2  (bases 1 to 4893)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol. 2014 Aug 8."
COMMENT     Sequence Label: pTE101
FEATURES             Location/Qualifiers
     source          1..4893
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      11..642
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      1105..1693
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1594..1613
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1849..2664)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1849..2664)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2572..2591
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2656..2675
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(2818..3966)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(4238..4609)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            4642..4751
                     /note="incP origin of transfer"
ORIGIN
        1 gaccctttcc gacgctcacc gggctggttg ccctcgccgc tgggctggcg gccgtctatg
       61 gccctgcaaa cgcgccagaa acgccgtcga agccgtgtgc gagacaccgc ggccgccggc
      121 gttgtggata cctcgcggaa aacttggccc tcactgacag atgaggggcg gacgttgaca
      181 cttgaggggc cgactcaccc ggcgcggcgt tgacagatga ggggcaggct cgatttcggc
      241 cggcgacgtg gagctggcca gcctcgcaaa tcggcgaaaa cgcctgattt tacgcgagtt
      301 tcccacagat gatgtggaca agcctgggga taagtgccct gcggtattga cacttgaggg
      361 gcgcgactac tgacagatga ggggcgcgat ccttgacact tgaggggcag agtgctgaca
      421 gatgaggggc gcacctattg acatttgagg ggctgtccac aggcagaaaa tccagcattt
      481 gcaagggttt ccgcccgttt ttcggccacc gctaacctgt cttttaacct gcttttaaac
      541 caatatttat aaaccttgtt tttaaccagg gctgcgccct gtgcgcgtga ccgcgcacgc
      601 cgaagggggg tgccccccct tctcgaaccc tcccggcccg ctaacgcggg cctcccatcc
      661 ccccaggggc tgcgcccctc ggccgcgaac ggcctcaccc caaaaatggc agccaagctg
      721 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
      781 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
      841 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
      901 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
      961 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     1021 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1081 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1141 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1201 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     1261 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1321 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1381 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1441 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     1501 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     1561 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     1621 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     1681 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     1741 ttttgctcac atggaattct gtacatctag aaataagaag gagatataat taatccatgg
     1801 caattgaagc ttagatcttg actagtcctg caggtaccta attcctgatt agaaaaactc
     1861 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg
     1921 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag
     1981 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc
     2041 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga
     2101 gaatggcaaa agcttatgca tttctttcca gacttgttca acaggccagc cattacgctc
     2161 gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag
     2221 acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg
     2281 caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac
     2341 ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg
     2401 gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat
     2461 ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc
     2521 atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc
     2581 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc tcgagcaaga
     2641 cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag
     2701 ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga
     2761 cacaacgtgg ctagcgctgg ctgctgaacc cccagccgga actgacccca caaggcccta
     2821 gcgtttgcaa tgcaccaggt catcattgac ccaggcgtgt tccaccaggc cgctgcctcg
     2881 caactcttcg caggcttcgc cgacctgctc gcgccacttc ttcacgcggg tggaatccga
     2941 tccgcacatg aggcggaagg tttccagctt gagcgggtac ggctcccggt gcgagctgaa
     3001 atagtcgaac atccgtcggg ccgtcggcga cagcttgcgg tacttctccc atatgaattt
     3061 cgtgtagtgg tcgccagcaa acagcacgac gatttcctcg tcgatcagga cctggcaacg
     3121 ggacgttttc ttgccacggt ccaggacgcg gaagcggtgc agcagcgaca ccgattccag
     3181 gtgcccaacg cggtcggacg tgaagcccat cgccgtcgcc tgtaggcgcg acaggcattc
     3241 ctcggccttc gtgtaatacc ggccattgat cgaccagccc aggtcctggc aaagctcgta
     3301 gaacgtgaag gtgatcggct cgccgatagg ggtgcgcttc gcgtactcca acacctgctg
     3361 ccacaccagt tcgtcatcgt cggcccgcag ctcgacgccg gtgtaggtga tcttcacgtc
     3421 cttgttgacg tggaaaatga ccttgttttg cagcgcctcg cgcgggattt tcttgttgcg
     3481 cgtggtgaac agggcagagc gggccgtgtc gtttggcatc gctcgcatcg tgtccggcca
     3541 cggcgcaata tcgaacaagg aaagctgcat ttccttgatc tgctgcttcg tgtgtttcag
     3601 caacgcggcc tgcttggcct cgctgacctg ttttgccagg tcctcgccgg cggtttttcg
     3661 cttcttggtc gtcatagttc ctcgcgtgtc gatggtcatc gacttcgcca aacctgccgc
     3721 ctcctgttcg agacgacgcg aacgctccac ggcggccgat ggcgcgggca gggcaggggg
     3781 agccagttgc acgctgtcgc gctcgatctt ggccgtagct tgctggacca tcgagccgac
     3841 ggactggaag gtttcgcggg gcgcacgcat gacggtgcgg cttgcgatgg tttcggcatc
     3901 ctcggcggaa aaccccgcgt cgatcagttc ttgcctgtat gccttccggt caaacgtccg
     3961 attcattcac cctccttgcg ggattgcccc gactcacgcc ggggcaatgt gcccttattc
     4021 ctgatttgac ccgcctggtg ccttggtgtc cagataatcc accttatcgg caatgaagtc
     4081 ggtcccgtag accgtctggc cgtccttctc gtacttggta ttccgaatct tgccctgcac
     4141 gaataccagc tccgcgaagt cgctcttctt gatggagcgc atggggacgt gcttggcaat
     4201 cacgcgcacc ccccggccgt tttagcggct aaaaaagtca tggctctgcc ctcgggcgga
     4261 ccacgcccat catgaccttg ccaagctcgt cctgcttctc ttcgatcttc gccagcaggg
     4321 cgaggatcgt ggcatcaccg aaccgcgccg tgcgcgggtc gtcggtgagc cagagtttca
     4381 gcaggccgcc caggcggccc aggtcgccat tgatgcgggc cagctcgcgg acgtgctcat
     4441 agtccacgac gcccgtgatt ttgtagccct ggccgacggc cagcaggtag gcctacaggc
     4501 tcatgccggc cgccgccgcc ttttcctcaa tcgctcttcg ttcgtctgga aggcagtaca
     4561 ccttgatagg tgggctgccc ttcctggttg gcttggtttc atcagccatc cgcttgccct
     4621 catctgttac gccggcggta gccggccagc ctcgcagagc aggattcccg ttgagcaccg
     4681 ccaggtgcga ataagggaca gtgaagaagg aacacccgct cgcgggtggg cctacttcac
     4741 ctatcctgcc cggctgacgc cgttggatac accaaggaaa gtctacacga accctttggc
     4801 aaaatcctgt atatcgtgcg aaaaaggatg gatataccga aaaaatcgct ataatgaccc
     4861 cgaagcaggg ttatgcagcg gaaaagatcc gtc
//
